Expression, Purification and Characterization of Recombinant Histidine-tagged L-asparaginase II | ||
| Egyptian Journal of Botany | ||
| Article 9, Volume 56, Issue 3, September 2016, Pages 707-722 PDF (374.54 K) | ||
| Document Type: Regular issue (Original Article) | ||
| DOI: 10.21608/ejbo.2016.2728 | ||
| Abstract | ||
| ESCHERICHIA coli has two L-asparaginase isozymes that have ..... been designated L-asparaginase I and L-asparaginase II. The amino acid sequences of both are rather dissimilar except for a few regions of significant homology. The sequence corresponding to the mature (ansB) was subcloned into pQE-30 expression vector and expressed in E. coli M15. The recombinant histidine-tagged (ansB) was purified to homogeneity by Ni–NTA affinity chromatography and displayed a single 36.0 kDa band on SDS-PAGE. Results revealed that the recombinant His-tagged L-ASNase II was expressed in an active form and its specific activity was estimated to be 286 U/mg. The optimum temperature was attained at 40°C. The enzyme was maximum at pH 7-8. Also the enzyme was stable at 40-50°C. The purified functional enzyme exhibited a specific activity at 286 U/mg and inhibited the growth of human myeloid leukemia cell line (HL-60) with IC50 value of 0.14±0.03 U/ml. | ||
| Keywords | ||
| Anti-tumor; Cytotoxicity; expression; His-tag; L-asparaginase; Leukemia; Purification; Recombinant | ||
|
Statistics Article View: 387 PDF Download: 788 |
||
